title: Bactericidal/permeability-increasing protein has endotoxin-neutralizing activity.
abstract: Neutrophil granules contain proteins important in host defense against bacterial pathogens. Granule proteins released from activated neutrophils facilitate opsonization, phagocytosis, tissue digestion, and antimicrobial activity. Three similar, if not identical, neutrophil proteins, bactericidal/permeability-increasing protein (BPI), 57,000 m.w. cationic antimicrobial protein, and bactericidal protein have been described that specifically kill gram negative bacteria. Since LPS is a structure common to all gram-negative bacteria, we investigated whether the microbicidal protein BPI affects biologic activity of LPS in vitro. Human neutrophils can be activated both in vitro and in vivo by LPS. Upon stimulation, surface expression of CR1 and CR3 increases markedly. Using flow microfluorimetry, we analyzed surface expression of CR1 and CR3 as a measure of neutrophil stimulation in response to LPS. CR up-regulation on neutrophils was TNF independent, suggesting direct LPS stimulation of neutrophils in this system. Purified BPI completely inhibited CR up-regulation on neutrophils stimulated with both rough and smooth LPS chemotypes at 1.8 to 3.6 nM (100 to 200 ng/ml). By comparison, the polypeptide antibiotic polymyxin B completely inhibited the same dose of LPS at 0.4 nM. The inhibitory activity of BPI appeared to be specific for LPS because neutrophil stimulation by formylated peptide or TNF was unaffected. The specificity of BPI for LPS was further demonstrated by inhibition of LPS activity in the limulus amebocyte lysate assay. Therefore, the role of BPI in infection may not be limited to its microbicidal activity, but it may also regulate the neutrophil response to LPS.
authors.affiliation: Invitron Corporation, Redwood City, CA 94063.
authors.forename: M N
authors.initials: MN
authors.lastname: Marra
authors.forename: C G
authors.initials: CG
authors.lastname: Wilde
authors.forename: J E
authors.initials: JE
authors.lastname: Griffith
authors.forename: J L
authors.initials: JL
authors.lastname: Snable
authors.forename: R W
authors.initials: RW
authors.lastname: Scott
mesh_terms: D023181:Antimicrobial Cationic Peptides; D001770:Blood Bactericidal Activity; D001798:Blood Proteins / Q000494:pharmacology*; D002463:Cell Membrane Permeability; D004731:Endotoxins / Q000037:antagonists & inhibitors*; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008033:Limulus Test; D008050:Lipid A / Q000037:antagonists & inhibitors; D008565:Membrane Proteins; D008970:Molecular Weight; D009504:Neutrophils / Q000502:physiology*; D011951:Receptors, Complement / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D023181:Antimicrobial Cationic Peptides; D001798:Blood Proteins; D004731:Endotoxins; D008050:Lipid A; D008565:Membrane Proteins; D011951:Receptors, Complement; D014409:Tumor Necrosis Factor-alpha; C021623:bactericidal permeability increasing protein